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Fully automatable two-dimensional hydrophilic interaction liquid chromatography-reversed phase liquid chromatography with online tandem mass spectrometry for shotgun proteomics
Yun Zhao1; Ricky P. W. Kong1; Guohui Li1,2; Maggie P. Y. Lam1; C. H. Law1; Simon M. Y. Lee2; Herman C. Lam1; Ivan K. Chu1
2012
Source PublicationJournal of Separation Science
ISSN16159306
Volume35Issue:14Pages:1755
Abstract

We have developed a fully automatable two-dimensional liquid chromatography platform for shotgun proteomics analyses based on the online coupling of hydrophilic interaction liquid chromatography (HILIC) - using a nonionic type of TSKgel Amide 80 at either pH 6.8 (neutral) or 2.7 (acidic) - with conventional low-pH reversed-phase chromatography. Online coupling of the neutral-pH HILIC and reversed phase chromatography systems outperformed the acidic HILIC-reversed phase chromatography combination, resulting in 18.4% (1914 versus 1617 nonredundant proteins) and 41.6% (12,989 versus 9172 unique peptides) increases in the number of identified peptides and proteins from duplicate analyses of Rat pheochromocytoma lysates. Armed with this optimized HILIC-reversed phase liquid chromatography platform, we identified 2554 nonredundant proteins from duplicate analyses of a Saccharomyces cerevisiae lysate, with the detected protein abundances spanning from approximately 41 to 106 copies per cell, which contained up to approximately 2092 different validated protein species with a dynamic range of concentrations of up to approximately 10 4. This present study establishes a fully automated platform as a promising methodology to enable online coupling of different hydrophilic HILIC and reversed phase chromatography systems, thereby expanding the repertoire of multidimensional liquid chromatography for shotgun proteomics.

KeywordHilic-rp Orthogonality Proteomics Two-dimensional Liquid Chromatography
DOI10.1002/jssc.201200054
URLView the original
Indexed BySCIE
Language英语
WOS Research AreaChemistry
WOS SubjectChemistry, Analytical
WOS IDWOS:000306493700008
The Source to ArticleScopus
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Cited Times [WOS]:27   [WOS Record]     [Related Records in WOS]
Document TypeJournal article
CollectionInstitute of Chinese Medical Sciences
Corresponding AuthorIvan K. Chu
Affiliation1.Department of Chemistry, The University of Hong Kong, Hong Kong, China
2.State Key Laboratory of Quality Research in Chinese Medicine and Institute of Chinese Medical Sciences, University of Macau, Taipa, Macao SAR, China
Recommended Citation
GB/T 7714
Yun Zhao,Ricky P. W. Kong,Guohui Li,et al. Fully automatable two-dimensional hydrophilic interaction liquid chromatography-reversed phase liquid chromatography with online tandem mass spectrometry for shotgun proteomics[J]. Journal of Separation Science,2012,35(14):1755.
APA Yun Zhao.,Ricky P. W. Kong.,Guohui Li.,Maggie P. Y. Lam.,C. H. Law.,...&Ivan K. Chu.(2012).Fully automatable two-dimensional hydrophilic interaction liquid chromatography-reversed phase liquid chromatography with online tandem mass spectrometry for shotgun proteomics.Journal of Separation Science,35(14),1755.
MLA Yun Zhao,et al."Fully automatable two-dimensional hydrophilic interaction liquid chromatography-reversed phase liquid chromatography with online tandem mass spectrometry for shotgun proteomics".Journal of Separation Science 35.14(2012):1755.
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